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[1]楊黎麗sup>,黃勝杰,李媚,等.溫陽補腎藥對抗骨髓間充質(zhì)干細胞凋亡的實驗研究[J].中醫(yī)正骨,2013,25(02):3-7.
　HUANG Sheng-jie*,YANG Li-li,LI Mei,et al.Mechanism of action of warming recuperating kidney medicine against the apoptosis of bone marrow stem cells[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2013,25(02):3-7.
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溫陽補腎藥對抗骨髓間充質(zhì)干細胞凋亡的實驗研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第25卷
期數(shù):
2013年02期
頁碼:
3-7
欄目:
基礎(chǔ)研究
出版日期:
2013-02-20

文章信息/Info

Title:
Mechanism of action of warming recuperating kidney medicine against the apoptosis of bone marrow stem cells
作者:
楊黎麗<sup>1; 黃勝杰2; 李媚1; 王和鳴1
1.福建中醫(yī)藥大學(xué),福建 福州 350003;
2.馬來西亞南方大學(xué),柔佛 新山 81300
Author(s):
HUANG Sheng-jie*; YANG Li-li; LI Mei; WANG He-ming.
*Fujian University of Traditional Chinese Medicine,Fuzhou 350003,Fujian,China
關(guān)鍵詞:
細胞凋亡 溫補腎陽 骨髓間充質(zhì)干細胞
Keywords:
Apoptosis;  WARMING RECUPERATING KIDNEY;  Bone marrow stem cell
摘要:
目的:探討溫陽補腎藥對骨髓間充質(zhì)干細胞凋亡的影響及作用機制。方法:建立白細胞介素1β誘導(dǎo)體外培養(yǎng)的SD大鼠骨髓間充質(zhì)干細胞凋亡體系,分別向其中加入巴戟天含藥血清(A組)、鹿角膠含藥血清(B組)、淫羊藿含藥血清(C組)、骨碎補含藥血清(D組)和空白血清(F組),E組不加入血清。各組細胞培養(yǎng)48 h后,采用RT-PCR法檢測bcl-2和Bax的mRNA表達量,采用電泳法檢測骨髓間充質(zhì)干細胞凋亡體系的建立,采用Western Blot法檢測bcl-2和Bax的蛋白表達量。結(jié)果:①骨髓間充質(zhì)干細胞表面標記結(jié)果。流式細胞儀檢測結(jié)果顯示CD34、CD45表達陰性,CD90表達陽性。②bcl-2和Bax的mRNA表達量。各組bcl-2mRNA表達量比較,差異有統(tǒng)計學(xué)意義(22.25±1.15,15.18±1.51,15.36±0.72,16.12±0.95,16.79±1.39,16.02±1.14; F=182.000,P=0.000)。A組bcl-2mRNA表達量高于B組、C組、D組、E組和F組(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000); E組高于B組(P=0.030)。各組BaxmRNA表達量比較,差異有統(tǒng)計學(xué)意義(19.17±0.67,15.37±1.02,18.18±0.23,17.37±0.72,28.36±0.93,23.52±1.26; F=28.422,P=0.000)。A組BaxmRNA表達量高于B組和D組(P=0.001,P=0.018),低于E組(P=0.001); B組低于C組、E組和F組(P=0.013,P=0.000,P=0.000),B組與D組比較,差異無統(tǒng)計學(xué)意義(P=0.052); D組低于F組(P=0.000); E組高于C組和F組(P=0.000,P=0.000)。③bcl-2DNA和BaxDNA電泳結(jié)果。電泳圖譜顯示,E組條帶與A組、B組、C組、D組有明顯差異。④bcl-2和Bax的蛋白表達量。各組bcl-2蛋白表達量比較,差異有統(tǒng)計學(xué)意義(11 526.18±697.38,20 096.88±953.73,16 784.88±504.36,14 856.65±546.90,8 549.29±313.63,9 004.79±399.12; F=341.740,P=0.000)。A組bcl-2蛋白表達量低于B組、C組和D組(P=0.000,P=0.000,P=0.000),高于E組和F組(P=0.000,P=0.000); B組高于C組、D組、E組和F組(P=0.000,P=0.000,P=0.000,P=0.000); C組高于D組、E組和F組(P=0.000,P=0.000,P=0.000); D組高于E組和F組(P=0.000,P=0.000)。各組Bax蛋白表達量比較,差異有統(tǒng)計學(xué)意義(12 435.09±1002.35,4 590.96±71.45,11 721.90±855.08,10 393.54±662.79,13 874.83±541.16,12 774.15±674.40; F=136.305,P=0.000)。A組Bax蛋白表達量高于B組和D組(P=0.000,P=0.039); B組低于C組、D組、E組和F組(P=0.000,P=0.000,P=0.000,P=0.000); C組低于E組(P=0.010); D組低于E組和F組(P=0.000,P=0.002)。結(jié)論:溫陽補腎藥含藥血清有對抗白細胞介素1β誘導(dǎo)的骨髓間充質(zhì)干細胞凋亡的作用,其機制主要是上調(diào)抑制凋亡基因Bcl-2的表達,下調(diào)促進凋亡基因Bax的表達,抑制其蛋白酶活性從而降低細胞凋亡率。
Abstract:
Objective:To explore the effect and mechanism of warming recuperating kidney medicine against the apoptosis of bone marrow stem cell(BMSC).Methods:The apoptosis system of SD rats BMSC cultured in vitro was established,and the apoptosis was induced by interleukin-1β(IL-1β),then the cells were respectively added with morinda root medicated serum(group A),deer-horn glue medicated serum(group B),epimedium herb medicated serum(group C),drynaria fortunei medicated serum(group D)and blank serum(group F),while group E without serum.After culturing 48 h for cells in each group,RT-PCR assay was used to detect mRNA expression levels of bcl-2 and Bax; electrophoresis method used to test the establishment of BMSC apoptosis system; and Western Blot method used to determine the protein expression levels of bcl-2 and Bax.Results:①The results of BMSC surface marker:the test results from flow cytometer were shown that CD34 and CD45 with negative expression,while CD90 with positive expression.②mRNA expression levels of bcl-2 and Bax:there was statistical difference in bcl-2mRNA expression level among all the groups(22.25±1.15,15.18±1.51,15.36±0.72,16.12±0.95,16.79±1.39,16.02±1.14; F=182.000,P=0.000).bcl-2mRNA expression level of group A was higher than that of group B,C,D,E and group F respectivrly(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000); group E was higher than group B(P=0.030).There was statistical difference in Bax mRNA expression level among all the groups(19.17±0.67,15.37±1.02,18.18±0.23,17.37±0.72,28.36±0.93,23.52±1.26; F=28.422,P=0.000).Bax mRNA expression level of group A was higher than that of group B and group D respectively(P=0.001,P=0.018),while lower than group E(P=0.001); group B was lower than group C,E and group F(P=0.013,P=0.000,P=0.000),while there was no statistical difference between group B and group D(P=0.052); group D was lower than group F(P=0.000); group E was higher than group C and group F respectively(P=0.000,P=0.000).③Electrophoresis results of bcl-2DNA and BaxDNA:there was significant difference in band between group E and group A,B,C and group D respectively through electrophoresis patterns.④Protein expression levels of bcl-2 and Bax:there was statistical difference in protein expression level of bcl-2 among all the groups(11 526.18±697.38,20 096.88±953.73,16 784.88±504.36,14 856.65±546.90,8 549.29±313.63,9 004.79±399.12; F=341.740,P=0.000).Protein expression level of bcl-2 of group A was lower than that of group B,C and group D respectively(P=0.000,P=0.000,P=0.000),while higher than that of group E and group F respectively(P=0.000,P=0.000); group B was higher than group C,D,E and group F respectively(P=0.000,P=0.000,P=0.000,P=0.000); group C was higher than group D,E and group F respectively(P=0.000,P=0.000,P=0.000); group D was higher than group E and group F respectively(P=0.000,P=0.000).There was statistical difference in protein expression level of Bax among all the groups(12 435.09±1 002.35,4 590.96±71.45,11 721.90±855.08,10 393.54±662.79,13 874.83±541.16,12 774.15±674.40; F=136.305,P=0.000).Protein expression level of Bax of group A was higher than that of group B and group D respectively(P=0.000,P=0.039); group B was lower than group C,D,E and group F respectively(P=0.000,P=0.000,P=0.000,P=0.000); group C was lower than group E(P=0.010); group D was lower than group E and group F respectively(P=0.000,P=0.002).Conclusion:The medicated serum of warming recuperating kidney medicine has the effect of inhibiting BMSC apoptosis induced by IL-1β,and its main mechanism is to inhibiting proteinase activity through up-regulation the expression of Bcl-2 while down-regulation the expression of Bax,then reduce the apoptosis rate.

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備注/Memo

備注/Memo:
基金項目:國家自然科學(xué)基金項目(30973763,81173283),教育部博士點基金-博導(dǎo)類項目(20093519110001)
通訊作者:王和鳴 E-mail:[email protected]
更新日期/Last Update: 2013-02-20